Bongekile Khoza ¹ Charles Byaruhanga ¹ Sekgota M. Makgabo ^1,2 Nkululeko Nyangiwe ³ Themba Mnisi ⁴ Samukelo Nxumalo ⁵ Marinda C. Oosthuizen ¹ Zamantungwa T. Mnisi ^1,6*

• ¹ Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa
• ² Department of Life and Consumer Sciences, University of South Africa, Florida, South Africa
• ³ Dohne Agriculture Development Institute, Stutterheim, South Africa
• ⁴ Other, Mbombela, South Africa
• ⁵ Other, Johannesburg, South Africa
• ⁶ Department of Biodiversity, Faculty of Science and Agriculture, University of Limpopo, Polokwane, South Africa

Ticks are obligate ectoparasites recognized worldwide as major vectors of several diseasecausing pathogens and are good indicators of disease distribution and epidemiology. Recent years has seen a growing concern regarding emerging and re-emerging of economically important tick-borne pathogens of livestock and humans worldwide. The overall objective of the study was to give an insight into current tick distribution and associated bacterial pathogens that may pose a threat to cattle in the sampled study sites. About 150 cattle were randomly selected from three study sites, Harrismith and Phuthaditjhaba in Free State Province and Bergville in KwaZulu Natal province, South Africa. Blood samples were collected from the cattle and DNA was subjected to the 16S rRNA gene microbiome sequencing using the circular consensus sequencing approach on the PacBio sequencing platform. Ticks were also collected from various predilection sites of the sampled animals. A total of eight tick species were identified and Rhipicephalus evertsi evertsi (79.4%) was the most abundant tick species, followed by R. appendiculatus (11.7%), R. afranicus (2.6%), R. simus (2.6%), Hyalomma rufipes (1.2%), R. decoloratus (1.0%), H. truncatum (0.7%) and R. microplus (0.7%). The bacterial microbiome sequence analysis revealed up to 16 phyla and 30 classes in the three study sites. Proteobacteria was the most dominant bacterial phyla with a relative abundance of 67.2% (Bergville), 73.8% (Harrismith) and 84.8% (Phuthaditjhaba), followed by Firmicutes at 9.6% (Phuthaditjhaba), 18.9% (Bergville) and 19.6% (Harrismith). The Chao 1 index estimator revealed significant differences in the α-diversity of microbial communities between three study sites. This study expands the knowledge on tick fauna and microbial communities in the three study sites.